Purpose: Physical activity questionnaire and measurement of cardiorespiratory fitness are two important methods of physical activity (PA) assessment. The purposes of this study were (1) to determine PA level using the PA questionnaire and cardiorespiratory fitness in undergraduate students (2) to detect gender differences in habitual physical activity. Materials and methods: 207 undergraduate students, aged 18 to 25 years participated in this study. 48.79% of the sample was female. PA was assessed with a PA questionnaire and classified into three PA groups (sedentary, active and very active). Cardiorespiratory fitness was measured in a 330 m outdoor track with one-mile walking test and predicted maximal oxygen consumption (VO2max) estimated by Kline's formula. Results: Moderate, vigorous and total PA scores were found higher in the both female and male very active groups when compared with sedentary and active groups (p<0.05). Similarly, the VO2max of the subjects in the very active group was higher than the other groups (p<0.05). Gender differences were found for moderate, vigorous activity scores and VO2max values (p<0.05). Vigorous activity scores and VO2max were higher in males than in females (p<0.05). Conclusion: The results of this study suggested that PA patterns of female and male students were different and VO2max associated with habitual PA.